Occurrence and ultrastructural localization of duck hepatitis B virus in the liver of ducks after experimental infection

Uchida, Tatsuo; Suzuki, Koyu; Esumi, Mariko; Arii, Masayuki; Oomura, Masashi; Shikata, Toshio

doi:10.1002/hep.1840070108

Cited by 9 publications

(7 citation statements)

References 17 publications

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“…The propagation of DHBV from positive to negative hepatocytes appeared to occur between Days 2 and 4 following inoculation of DHBV to 0-day-old ducklings (15). In contrast, propagation of DHBV secreted by positive cells to negative cells did not occur under cultured conditions because cultured hepatocytes with DHBV infection were individually scattered among the negative cells, and positive cells did not increase with time in days of culture.…”

Section: Resultsmentioning

confidence: 78%

“…As seen in cultivation of DHBV-positive hepatocytes infected in uiuo, cultured hepatocytes originally have the potential to continuously secrete viruses even after spindle-shape alteration, once they had been infected with DHBV (TJchida, T., unpublished observations). The propagation of DHBV from positive to negative hepatocytes appeared to occur between Days 2 and 4 following inoculation of DHBV to 0-day-old ducklings (15). In contrast, propagation of DHBV secreted by positive cells to negative cells did not occur under cultured conditions because cultured hepatocytes with DHBV infection were individually scattered among the negative cells, and positive cells did not increase with time in days of culture.…”

Section: Discussionmentioning

confidence: 89%

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In Vitro transmission of duck hepatitis B virus to primary duck hepatocyte cultures

et al. 1988

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An attempt was made to infect primary duck hepatocyte cultures with duck hepatitis B virus in vitro in order to clarify the biology of hepatitis B virus. Livers of ducklings, 0 to 17 days posthatch, without viremia were digested ex situ by perfusion of collagenase solution through the portal or hepatic vein. Homogeneous hepatocyte suspensions were seeded into plastic dishes in L-15 medium containing 10(-8) M insulin, 2 X 10(-8) M glucagon and 10(-8) M dexamethasone and were subsequently inoculated with sufficient numbers of duck hepatitis B virus. As a result, duck hepatitis B virus multiplication started weakly on Day 2, gradually increased and reached the maximum level approximately on Day 10 postinoculation. Viral replication was revealed by duck hepatitis B virus DNA in the cell pellet and in the culture medium and duck hepatitis B virus DNA-specific transcripts in the cell pellet. Immunostaining demonstrated duck hepatitis B virus core antigen in approximately 10% of cultured hepatocytes, and an increase in numbers of positive cells was not observed with time for up to 18 days of culture. Viral particles were found within the endoplasmic reticulum, and the inoculation of culture medium provoked viremia in the ducklings. The age of ducklings did not influence the numbers of cells infected. The in vitro infection system was similar to the in vivo one; however, the former seemed to be age-independent and to allow replication of duck hepatitis B virus in the limited number of hepatocytes.

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Section: Resultsmentioning

confidence: 78%

Section: Discussionmentioning

confidence: 89%

In Vitro transmission of duck hepatitis B virus to primary duck hepatocyte cultures

et al. 1988

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“…DHBV-positive serum was originally provided by T. Uchida, Nihon University School of Medicine, Tokyo, Japan (22). Ducklings were injected intravenously with 50 ,ul of the original DHBV-positive serum.…”

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confidence: 99%

Comparative activities of several nucleoside analogs against duck hepatitis B virus in vitro

Yokota

Konno

Chonan

et al. 1990

Antimicrob Agents Chemother

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Duck hepatitis B virus (DHBV) replication in primary duck hepatocytes was monitored by examining the synthesis of both DHBV DNA and DHBV core antigen. Several nucleoside analogs which were previously shown to inhibit the replication of DNA viruses (i.e., herpesviruses) and retroviruses were examined for their inhibitory effects on the synthesis of DHBV core antigen in primary duck hepatocytes. (S)-9-(3-Hydroxy-2-phosphonylmethoxypropyl)adenine [(S)-HPMPA], 9-(2-phosphonylmethoxyethyl)-2,6-diaminopurine, 2',3'-dideoxyadenosine, and 2',3'-dideoxycytidine inhibited DHBV core antigen synthesis at concentrations that were significantly lower than those found to be toxic to the primary hepatocytes. Of all the compounds tested, (S)-HPMPA showed the lowest 50% effective concentration (0.5 ,ug/ml). The selectivity index or ratio of the 50% cytotoxic concentration to the 50% effective concentration of (S)-HPMPA was greater than 300. (S)-HPMPA not only inhibited DHBV core antigen but also DHBV DNA synthesis in DHBV-infected hepatocytes.Human hepatitis B virus (HBV) is a causative agent of both an acute and chronic form of hepatitis. More than 200 million people worldwide are estimated to be chronic carriers of HBV. HBV belongs to the family Hepadnaviridae, which also includes woodchuck hepatitis virus, ground squirrel hepatitis virus, tree squirrel hepatitis virus, and duck hepatitis B virus (DHBV) (3,(7)(8)(9)(10)(11)(12)(13)(14)19). All members of the hepadnavirus family have a number of characteristics in common such as morphological appearance, antigenic makeup, and DNA size and structure (3,(7)(8)(9)(10)(11)(12)(13)(14)19). Also, the pathological findings following infection with these viruses are quite similar (8,20). The replication of the hepadnaviruses (4, 10) is dependent on the reverse transcription of an RNA intermediate. This results in the formation of a minusstranded DNA, which then serves as a template for the synthesis of plus-stranded DNA.DHBV is able to propagate in primary duck hepatocytes in vitro. Tuttleman et al. (21) reported that upon infection of primary duck hepatocytes with DHBV, viral DNA is synthesized, virus-specific proteins are produced, and infectious virus is released into the culture medium. Using immunofluorescence, they demonstrated the production of both core and surface DHBV antigens in primary duck hepatocytes that were infected with DHBV. This in vitro system should prove suitable to evaluate the effects of antiviral drugs against DHBV. In this study, we examined a wide variety of such drugs that belong to the class of the purine and pyrimidine nucleoside analogs for their inhibitory effects on replication of DHBV in primary duck hepatocytes in vitro. was from ICN Nutritional Biochemicals, Cleveland, Ohio; 2',3'-dideoxy-2',3'-didehydrothymidine (D4T), 2',3'-dideoxyuridine (DDU), 2',3'-dideoxyadenosine (DDA), 2',3'-dideoxycytidine (DDC), 3'-deoxyarabinofuranosylcytidine (D-ara-C), 3'-azido-2',3'-dideoxythymidine (AZT), 3'-azido-2',3'-dideoxyuridine (AZU), acyclovir [9-(2-hydroxyeth...

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“…8'11'12 Ducks have therefore been used as experimental animals for the study of HBV, since they are inexpensive and available in large numbers. [13][14][15] Studies of the pathogenesis and treatment of liver diseases due to HBV have moved towards virology, as the significance of various antigens and antibodies relating to HBV has been demonstrated. 1,16 Interferon, for example, has been used clinically and it is anticipated that it will have anti-viral effects against hepatitis C virus (HCV), in addition to showing such effects against HBV.…”

Section: Inoculation Of Medium To 1-day-old Duckmentioning

confidence: 99%

Effects of acyclovir and vidarabin 5′-monophosphate on anti-duck hepatitis B virus in an in vitro culture system

Motegi¹

1995

J Gastroenterol

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Various anti-viral agents, e.g., interferon, have recently been used for the treatment of viral hepatitis. In the present study, duck hepatitis B virus (DHBV) was cultured in vitro and the anti-DHBV effects of acyclovir (ACV) and vidarabin 5'-monophosphate (VMP) were studied. The portal perfusion method was applied to the livers of 7-day-old white ducks weighing 100 g, bred in Japan, and hepatocytes were infected with DHBV in vitro. Duck hepatocytes infected with DHBV were cultured in medium containing ACV or VMP, and the anti-DHBV effects of these drugs were assessed by determining DHBV-DNA and duck hepatitis B surface antigen in the medium. Both ACV and VMP had anti-DHBV effects when used immediately after infection; however, both drugs were ineffective in hepatocytes obtained from a DHBV carrier duck. In conclusion, the anti-DHBV effects of these drugs were very limited. However, this culture system appears to be useful for studies of hepatitis virus and anti-viral drugs.

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Occurrence and ultrastructural localization of duck hepatitis B virus in the liver of ducks after experimental infection

Cited by 9 publications

References 17 publications

In Vitro transmission of duck hepatitis B virus to primary duck hepatocyte cultures

In Vitro transmission of duck hepatitis B virus to primary duck hepatocyte cultures

Comparative activities of several nucleoside analogs against duck hepatitis B virus in vitro

Effects of acyclovir and vidarabin 5′-monophosphate on anti-duck hepatitis B virus in an in vitro culture system

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